Identification and cloning of the bacterial nodulation specificity gene in the Sinorhizobium meliloti – Medicago laciniata symbiosis

2002 ◽  
Vol 48 (9) ◽  
pp. 765-771 ◽  
Author(s):  
L R Barran ◽  
E S.P Bromfield ◽  
D C.W Brown
Keyword(s):  
Nucleotide Sequence ◽  
Medicago Sativa ◽  
Sinorhizobium Meliloti ◽  
Nucleotide Sequencing ◽  
Restricted Range ◽  
Tn5 Mutagenesis ◽  
Nodc Gene ◽  
Dna Fragment ◽  
Nodulation Specificity ◽  
Practical Implications

Medicago laciniata (cut-leaf medic) is an annual medic that is highly nodulation specific, nodulating only with a restricted range of Sinorhizobium meliloti, e.g., strain 102L4, but not with most strains that nodulate Medicago sativa (alfalfa), e.g., strains RCR2011 and Rm41. Our aim was to identify and clone the S. meliloti 102L4 gene implicated in the specific nodulation of M. laciniata and to characterize the adjacent nodulation (nod) region. An 11-kb EcoRI DNA fragment from S. meliloti 102L4 was shown to complement strain RCR2011 for nodulation of M. laciniata. Nucleotide sequencing revealed that this fragment contained nodABCIJ genes whose overall arrangement was similar to those found in strains RCR2011 and Rm41, which do not nodulate M. laciniata. Data for Tn5 mutagenesis of the nodABCIJ region of strain 102L4 suggested that the nodC gene was involved in the specific nodulation of M. laciniata. Tn5 insertions in the nodIJ genes gave mutants with nodulation delay phenotypes on both M. laciniata and M. sativa. Only subclones of the 11-kb DNA fragment containing a functional nodC gene from strain 102L4 were able to complement strain RCR2011 for nodulation of M. laciniata. The practical implications of these findings are discussed in the context of the development of a specific M. sativa – S. meliloti combination that excludes competition for nodulation by bacterial competitors resident in soil.Key words: Sinorhizobium meliloti, Medicago laciniata, nodulation specificity,nod gene, nucleotide sequence.

Sequencing Analysis and Phylogenetic Tree of HPV Isolated from Breast Cancer Patients at Thi-Qar Province/Iraq

2020 ◽  
pp. 37-40
Keyword(s):  
Nucleotide Sequence ◽  
Phylogenetic Tree ◽  
Evolutionary Relationship ◽  
Nucleotide Sequencing ◽  
Sequencing Analysis ◽  
Breast Cancer Patients ◽  
The North ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
History Of

Genetic variety examination has demonstrated fundamental to the understanding of the epidemiological and developmental history of Papillomavirus (HPV), for the development of accurate diagnostic tests and for efficient vaccine design. The HPV nucleotide diversity has been investigated widely among high-risk HPV types. To make the nucleotide sequence of HPV and do the virus database in Thi-Qar province, and compare sequences of our isolates with previously described isolates from around the world and then draw its phylogenetic tree, this study done. A total of 6 breast formalin-fixed paraffin-embedded (FFPE) of the female patients were included in the study, divided as 4 FFPE malignant tumor and 2 FFPE of benign tumor. The PCR technique was implemented to detect the presence of HPV in breast tissue, and the real-time PCR used to determinant HPV genotypes, then determined a complete nucleotide sequence of HPV of L1 capsid gene, and draw its phylogenetic tree. The nucleotide sequencing finding detects a number of substitution mutation (SNPs) in (L1) gene, which have not been designated before, were identified once in this study population, and revealed that the HPV16 strains have the evolutionary relationship with the South African race, while, the HPV33 and HPV6 showing the evolutionary association with the North American and East Asian race, respectively.

scholarly journals Molecular analysis of enterolysin a and entL gene cluster from natural isolate Enterococcus faecalis BGPT1-10P

Genetika ◽  
2013 ◽  
Vol 45 (2) ◽  
pp. 479-492 ◽  
Author(s):  
Katarina Veljovic ◽  
Amarela Terzic-Vidojevic ◽  
Maja Tolinacki ◽  
Milan Kojic ◽  
Ljubisa Topisirovic
Keyword(s):  
Functional Analysis ◽  
Nucleotide Sequence ◽  
Enterococcus Faecalis ◽  
Broad Spectrum ◽  
Candida Species ◽  
Genetic Information ◽  
Natural Isolate ◽  
Gene Encoding ◽  
Dna Fragment ◽  
Virulent Factor

Strain Enterococcus faecalis BGPT1-10P was isolated from artisanal semi-hard homemade cheese from Stara Planina, Serbia. Results showed that BGPT1-10P synthesized a heat labile bacteriocin with a broad spectrum of activity, including Listeria and Candida species. Further analysis revealed that synthesized bacteriocin is enterolysin A. Moreover, the entL gene encoding enterolysin A was found to be located on the chromosome. The entL gene was cloned and sequenced. Analysis of nucleotide sequence showed that the entL gene in natural isolate En. faecalis BGPT1-10P is identical to that of the entL gene described previously in En. faecalis LMG 2333. Within the cloned DNA fragment containing the entL gene, four ORFs were detected. One of them was identified as the scpE gene, which encodes a virulent factor staphopain peptidase. Functional analysis of the entL gene showed that the complete genetic information necessary for the synthesis of enterolysin A were directly linked solely to it. Strain BGPT1-10P also synthesized gelatinase and citolysin, and contained a set of virulent factors. In addition, BGPT1-10P carries the ermB and tetM genes conferring the resistance to erythromycin and tetracycline, respectively.

scholarly journals H2O2 Is Required for Optimal Establishment of the Medicago sativa/Sinorhizobium meliloti Symbiosis

2007 ◽  
Vol 189 (23) ◽  
pp. 8741-8745 ◽  
Author(s):  
Alexandre Jamet ◽  
Karine Mandon ◽  
Alain Puppo ◽  
Didier Hérouart
Keyword(s):  
Medicago Sativa ◽  
Plant Cell ◽  
Sinorhizobium Meliloti ◽  
Root Hairs ◽  
Symbiotic Interaction ◽  
Infection Threads ◽  
Cell Layers

ABSTRACT The symbiotic interaction between Medicago sativa and Sinorhizobium meliloti RmkatB ++ overexpressing the housekeeping catalase katB is delayed, and this delay is combined with an enlargement of infection threads. This result provides evidence that H2O2 is required for optimal progression of infection threads through the root hairs and plant cell layers.

scholarly journals The Sinorhizobium meliloti ABC Transporter Cho Is Highly Specific for Choline and Expressed in Bacteroids from Medicago sativa Nodules

2004 ◽  
Vol 186 (18) ◽  
pp. 5988-5996 ◽  
Author(s):  
Laurence Dupont ◽  
Isabelle Garcia ◽  
Marie-Christine Poggi ◽  
Geneviève Alloing ◽  
Karine Mandon ◽  
...  
Keyword(s):  
Medicago Sativa ◽  
Abc Transporter ◽  
Sinorhizobium Meliloti ◽  
Lipid Membrane ◽  
Site Directed Mutagenesis ◽  
Choline Uptake ◽  
Uptake System ◽  
Choline Transport ◽  
High Affinity ◽  
Uptake Activity

ABSTRACT In Sinorhizobium meliloti, choline is the direct precursor of phosphatidylcholine, a major lipid membrane component in the Rhizobiaceae family, and glycine betaine, an important osmoprotectant. Moreover, choline is an efficient energy source which supports growth. Using a PCR strategy, we identified three chromosomal genes (choXWV) which encode components of an ABC transporter: ChoX (binding protein), ChoW (permease), and ChoV (ATPase). Whereas the best homology scores were obtained with components of betaine ProU-like systems, Cho is not involved in betaine transport. Site-directed mutagenesis of choX strongly reduced (60 to 75%) the choline uptake activity, and purification of ChoX, together with analysis of the ligand-binding specificity, showed that ChoX binds choline with a high affinity (K D , 2.7 μM) and acetylcholine with a low affinity (K D , 145 μM) but binds none of the betaines. Uptake competition experiments also revealed that ectoine, various betaines, and choline derivatives were not effective competitors for Cho-mediated choline transport. Thus, Cho is a highly specific high-affinity choline transporter. Choline transport activity and ChoX expression were induced by choline but not by salt stress. Western blotting experiments with antibodies raised against ChoX demonstrated the presence of ChoX in bacteroids isolated from nitrogen-fixing nodules obtained from Medicago sativa roots. The choX mutation did not have an effect on growth under standard conditions, and neither Nod nor Fix phenotypes were impaired in the mutant, suggesting that the remaining choline uptake system(s) still present in the mutant strain can compensate for the lack of Cho transporter.

Nucleotide Sequence of a Cloned Murine Leukemia Virus DNA Fragment

1980 ◽  
Vol 44 (0) ◽  
pp. 1275-1279
Author(s):  
J. G. Sutcliffe ◽  
T. M. Shinnick ◽  
R. A. Lerner ◽  
P. Johnson ◽  
I. M. Verma
Keyword(s):  
Nucleotide Sequence ◽  
Murine Leukemia Virus ◽  
Leukemia Virus ◽  
Dna Fragment ◽  
Murine Leukemia
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